Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China; Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Nanjing, 210029, China; The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Institute of Translational Medicine, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211816, China.
J Ethnopharmacol. 2020 May 23;254:112737. doi: 10.1016/j.jep.2020.112737. Epub 2020 Mar 5.
Tongnao Decoction (TND) is a Chinese decoction approved and used in Jiangsu Province Hospital for the treatment of ischemic stroke. It shows conclusive efficiency in the improvement of neurologic impairment and activities of daily living of the patients.
Recently, angiogenesis has been recognized as a potential therapeutic strategy for treating cerebral ischemia. This study was aimed to provide comprehensive evidence for the pro-angiogenic effect of TND and characterize the underlying mechanism.
We firstly established the chemical fingerprinting of TND. Then, the in vitro pro-angiogenic activities of TND were tested on human umbilical vein endothelial cells (HUVECs) through cell viability, wound healing and tube formation assays. The in vivo pro-angiogenic effects were evaluated on transgenic zebrafish embryos [Tg (fli-1: EGFP)] through the formation of intersegmental vessels (ISVs), subintestinal vessels (SIVs) and central arteries (CtAs). Lastly, the potential mechanisms of TND were analyzed by a blocking assay with eight pathways-specific kinase inhibitors.
TND promoted the proliferation, migration and tube formation of HUVECs. TND also rescued the impairment of ISVs, SIVs and CtAs caused by VRI in a dose-dependent manner in zebrafish embryos. TND could activate vascular endothelial growth factor receptor-2 (VEGFR-2), phosphoinositide 3-kinase (PI3K) - protein kinase B (Akt) and Raf - mitogen-activated protein kinase1/2 (MEK1/2) - extracellular regulated kinase 1/2 (ERK1/2) signaling pathways.
Our study firstly demonstrated the pro-angiogenic activities of TND. Our work provided evidences for the clinical usage of TND in restoring neurovascular function through promoting angiogenesis in the ischemic cerebral microvascular.
通脑汤（TND）是一种中药方剂，已在江苏省中医院获得批准并用于治疗缺血性中风。它在改善患者神经功能缺损和日常生活活动能力方面显示出确切的疗效。
最近，血管生成已被认为是治疗脑缺血的一种潜在治疗策略。本研究旨在为 TND 的促血管生成作用提供全面的证据，并探讨其潜在的作用机制。
我们首先建立了 TND 的化学指纹图谱。然后，通过细胞活力、划痕愈合和管形成实验，在人脐静脉内皮细胞（HUVECs）上测试 TND 的体外促血管生成活性。通过转(transgenic)斑马鱼胚胎[Tg(fli-1: EGFP)]的节间血管（ISVs）、肠下血管（SIVs）和中央动脉（CtAs）形成来评估 TND 的体内促血管生成作用。最后，通过使用八种通路特异性激酶抑制剂的阻断实验分析 TND 的潜在作用机制。
TND 促进了 HUVECs 的增殖、迁移和管形成。TND 还以剂量依赖性方式挽救了 VRI 引起的斑马鱼胚胎中 ISVs、SIVs 和 CtAs 的损伤。TND 可以激活血管内皮生长因子受体-2（VEGFR-2）、磷脂酰肌醇 3-激酶（PI3K）-蛋白激酶 B（Akt）和 Raf-丝裂原激活蛋白激酶 1/2（MEK1/2）-细胞外调节激酶 1/2（ERK1/2）信号通路。
本研究首次证明了 TND 的促血管生成活性。我们的工作为 TND 通过促进缺血性脑血管中的血管生成来恢复神经血管功能的临床应用提供了证据。
